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Xenopus as a model system for the study of GOLPH2/GP73 function: Xenopus GOLPH2 is required for pronephros development.

Li L, Wen L, Gong Y, Mei G, Liu J, Chen Y, Peng T - PLoS ONE (2012)

Bottom Line: The morpholino-mediated knockdown of golph2 results in edema formation.Expression patterns of the transcription factors WT1, Pax2, Pax8, Lim1, GATA3, and HNF1β are also examined in the golph2 knockdown embryos, the expression of WT1 is increased in the glomus and expanded laterally in the pronephric region.We conclude that the deletion of golph2 causes an increase in the expression of WT1, which may promote glomus formation and inhibit pronephric tubule differentiation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

ABSTRACT
GOLPH2 is a highly conserved protein. It is upregulated in a number of tumors and is being considered as an emerging biomarker for related diseases. However, the function of GOLPH2 remains unknown. The Xenopus model is used to study the function of human proteins. We describe the isolation and characterization of Xenopus golph2, which dimerizes and localizes to the Golgi in a manner similar to human GOLPH2. Xenopus golph2 is expressed in the pronephros during early development. The morpholino-mediated knockdown of golph2 results in edema formation. Additionally, Nephrin expression is enhanced in the glomus, and the expression of pronephric marker genes, such as atp1b1, ClC-K, NKCC2, and NBC1, is diminished in the tubules and duct. Expression patterns of the transcription factors WT1, Pax2, Pax8, Lim1, GATA3, and HNF1β are also examined in the golph2 knockdown embryos, the expression of WT1 is increased in the glomus and expanded laterally in the pronephric region. We conclude that the deletion of golph2 causes an increase in the expression of WT1, which may promote glomus formation and inhibit pronephric tubule differentiation.

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Xenopus golph2 localizes to the Golgi. A.Perinuclear localization of endogenous golph2 in the pronephros of stage 46/47 embryos. B. Pronephric tubule. C–E. Endogenous golph2 co-localizes with hGalNAcT2-EGFP in animal cap cells. The mRNA of hGalNAcT2-EGFP was injected into the animal pole of the fertilized eggs (C, green); golph2 was detected using the antibody 10F12 followed by Cy3-conjugated secondary antibody (D, red); merge (E). F–H. Endogenous golph2 co-localizes with xGalNAcT2-EGFP in animal cap cells. xGalNAcT2-EGFP (F, green); golph2 (G, red); merge (H). Bar = 10 µm.
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pone-0038939-g002: Xenopus golph2 localizes to the Golgi. A.Perinuclear localization of endogenous golph2 in the pronephros of stage 46/47 embryos. B. Pronephric tubule. C–E. Endogenous golph2 co-localizes with hGalNAcT2-EGFP in animal cap cells. The mRNA of hGalNAcT2-EGFP was injected into the animal pole of the fertilized eggs (C, green); golph2 was detected using the antibody 10F12 followed by Cy3-conjugated secondary antibody (D, red); merge (E). F–H. Endogenous golph2 co-localizes with xGalNAcT2-EGFP in animal cap cells. xGalNAcT2-EGFP (F, green); golph2 (G, red); merge (H). Bar = 10 µm.

Mentions: Previously, we demonstrated that the Golgi retention of GOLPH2 is determined by the TMD [7]. The TMD of golph2 is highly conserved with GOLPH2, thereby suggesting that golph2 may localize to the Golgi. As shown in Fig. 2A, perinuclear localization of endogenous golph2 was observed in stage 46/47 embryonic pronephros. It has been shown that N-terminal of human hGalNACT2 localized in the Golgi of Xenopus A6 cell lines [19]. We use the EGPF-fusion protein of hGalNACT2 and the Xenopus xGalNACT2 as Golgi markers. We detected that endogenous golph2 co-localized with both of the markers in Xenopus animal cap cells (Fig. 2C–2H).


Xenopus as a model system for the study of GOLPH2/GP73 function: Xenopus GOLPH2 is required for pronephros development.

Li L, Wen L, Gong Y, Mei G, Liu J, Chen Y, Peng T - PLoS ONE (2012)

Xenopus golph2 localizes to the Golgi. A.Perinuclear localization of endogenous golph2 in the pronephros of stage 46/47 embryos. B. Pronephric tubule. C–E. Endogenous golph2 co-localizes with hGalNAcT2-EGFP in animal cap cells. The mRNA of hGalNAcT2-EGFP was injected into the animal pole of the fertilized eggs (C, green); golph2 was detected using the antibody 10F12 followed by Cy3-conjugated secondary antibody (D, red); merge (E). F–H. Endogenous golph2 co-localizes with xGalNAcT2-EGFP in animal cap cells. xGalNAcT2-EGFP (F, green); golph2 (G, red); merge (H). Bar = 10 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3375297&req=5

pone-0038939-g002: Xenopus golph2 localizes to the Golgi. A.Perinuclear localization of endogenous golph2 in the pronephros of stage 46/47 embryos. B. Pronephric tubule. C–E. Endogenous golph2 co-localizes with hGalNAcT2-EGFP in animal cap cells. The mRNA of hGalNAcT2-EGFP was injected into the animal pole of the fertilized eggs (C, green); golph2 was detected using the antibody 10F12 followed by Cy3-conjugated secondary antibody (D, red); merge (E). F–H. Endogenous golph2 co-localizes with xGalNAcT2-EGFP in animal cap cells. xGalNAcT2-EGFP (F, green); golph2 (G, red); merge (H). Bar = 10 µm.
Mentions: Previously, we demonstrated that the Golgi retention of GOLPH2 is determined by the TMD [7]. The TMD of golph2 is highly conserved with GOLPH2, thereby suggesting that golph2 may localize to the Golgi. As shown in Fig. 2A, perinuclear localization of endogenous golph2 was observed in stage 46/47 embryonic pronephros. It has been shown that N-terminal of human hGalNACT2 localized in the Golgi of Xenopus A6 cell lines [19]. We use the EGPF-fusion protein of hGalNACT2 and the Xenopus xGalNACT2 as Golgi markers. We detected that endogenous golph2 co-localized with both of the markers in Xenopus animal cap cells (Fig. 2C–2H).

Bottom Line: The morpholino-mediated knockdown of golph2 results in edema formation.Expression patterns of the transcription factors WT1, Pax2, Pax8, Lim1, GATA3, and HNF1β are also examined in the golph2 knockdown embryos, the expression of WT1 is increased in the glomus and expanded laterally in the pronephric region.We conclude that the deletion of golph2 causes an increase in the expression of WT1, which may promote glomus formation and inhibit pronephric tubule differentiation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

ABSTRACT
GOLPH2 is a highly conserved protein. It is upregulated in a number of tumors and is being considered as an emerging biomarker for related diseases. However, the function of GOLPH2 remains unknown. The Xenopus model is used to study the function of human proteins. We describe the isolation and characterization of Xenopus golph2, which dimerizes and localizes to the Golgi in a manner similar to human GOLPH2. Xenopus golph2 is expressed in the pronephros during early development. The morpholino-mediated knockdown of golph2 results in edema formation. Additionally, Nephrin expression is enhanced in the glomus, and the expression of pronephric marker genes, such as atp1b1, ClC-K, NKCC2, and NBC1, is diminished in the tubules and duct. Expression patterns of the transcription factors WT1, Pax2, Pax8, Lim1, GATA3, and HNF1β are also examined in the golph2 knockdown embryos, the expression of WT1 is increased in the glomus and expanded laterally in the pronephric region. We conclude that the deletion of golph2 causes an increase in the expression of WT1, which may promote glomus formation and inhibit pronephric tubule differentiation.

Show MeSH
Related in: MedlinePlus